Cell Painting unravels insecticidal modes of action on Spodoptera frugiperda insect cells.

The "Cell Painting" technology utilizes multiplexed fluorescent staining of various cell organelles, to produce high-content microscopy images of cells for multidimensional phenotype assessment. The phenotypic profiles extracted from those images can be analyzed upon perturbations with biologically active molecules to annotate the mode of action or biological activity by comparison with reference profiles of already known mechanisms of action, ultimately enabling the determination of on-target and off-target effects. This approach is already described in various human cell cultures, the most commonly used being the U2OS cell line, yet allows broad applications in additional areas of chemical-biological research. Here we describe for the first time the application and adaptation of Cell Painting to an insect cell line, the Sf9 cells from Spodoptera frugiperda. By adjusting image acquisition and analysis models, specific phenotypic profiles were obtained in a dose-dependent manner for 20 reference compounds, including representatives for the most relevant insecticidal modes of action categories (nerve & muscle, respiration and growth & development). Through a dimensionality-reduction method, both calculations of phenotypic half maximal inhibition concentration (IC50) values as well as similarity analysis of the obtained profiles by hierarchical clustering were performed. By Cell Painting effects on the phenotype could be obtained at higher sensitivity than in other assay formats, such as cytotoxicity assessments. More importantly, these analyses provide insight into mechanistic determinants of biological activity. Compounds with similar modes of action showed a high degree of proximity in a hierarchical clustering analysis while being distinct from actives with an unrelated mode of action. In essence, we provide strong evidence on the impact of Cell Painting mechanistic understanding of insecticides with regards to determinants of efficacy and safety utilizing an insect cell model system.

Bioresorbable scaffolds for the treatment of in-stent restenosis.

Percutaneous coronary intervention with bare-metal stents or drug-eluting stents can decrease clinical event rates compared with simple balloon angioplasty. However, stent implantation is often associated with subsequent restenosis. Bioresorbable coronary scaffolds provide short-term vessel scaffolding with drug delivery capability and are designed to avoid the long-term limitations of metallic stents such as late stent thrombosis and in-stent restenosis. To the best of our knowledge, we report the first case series of successful treatment of in-stent restenosis using bioresorbable scaffolds.

Reproducibility of aortic annulus measurements by computed tomography.

OBJECTIVES: To evaluate a systematic approach for measurement of aortic annulus dimensions by cardiac computed tomography. METHODS: CT data sets of 64 patients were evaluated. An oblique cross-section aligned with the aortic root was created by systematically identifying the caudal insertion points of the three aortic cusps and sequentially aligning them in a double oblique plane. Aortic annulus dimensions, distances of coronary ostia and a suitable fluoroscopic projection angle were independently determined by two observers. RESULTS: Interobserver intraclass correlation coefficients (ICC) for aortic annulus diameters were excellent (ICC 0.89-0.93). Agreement for prosthesis size selection was excellent (ĸ = 0.86 for mean, ĸ = 0.84 for area-derived and ĸ = 0.91 for circumference-derived diameter). Mean distances of the left/right coronary ostium were 13.4 ± 2.4/14.4 ± 2.8 mm for observer 1 and 13.2 ± 2.7/13.5 ± 3.2 mm for observer 2 (p = 0.30 and p = 0.0001, respectively; ICC 0.76/0.77 for left/right coronary artery). A difference of less than 10° for fluoroscopic projection angle was achieved in 84.3% of patients. CONCLUSIONS: A systematic approach to generate a double oblique imaging plane exactly aligned with the aortic annulus demonstrates high interobserver and intraobserver agreements for derived measurements which are not influenced by aortic root calcification. KEY POINTS: • Systematic approach to generate a double oblique imaging plane for TAVI evaluation. • This method is straightforward and software independent. • An approach with high reproducibility, not influenced by aortic root calcification.

Influence of renal sympathetic denervation on quality of life.

BACKGROUND: Therapy-resistant arterial hypertension causing psychosocial stress and is associated with cardiovascular morbidity and mortality. The aim of the study was to evaluate the effect on quality of life (QoL) in patients with resistant hypertension undergoing renal sympathetic denervation (RSD). METHODS AND RESULTS: We analyzed responses to the SF-36 Quality of Life Questionnaire provided by patients with resistant arterial hypertension after RSD. Thirty consecutive patients from 2 centers were included in this study, from October 2011 until February 2012. The phone interview was performed after the 3-month follow-up. A significant reduction (26 ± 13.5 mmHg) in systolic blood pressure (BP) was detected at the 3-month follow-up (142.0 ± 15.1 mmHg vs 168.0 ± 13.7 mmHg; P < 0.001). Seventy-five percent of the patients indicated that their health situation was a lot better (better, 21%; equal to, 4%) 3 months after RSD compared to the time before the therapeutic procedure. Furthermore, the majority of patients felt full of pep (always, 29%; mostly, 58%; quite often, 8%; sometimes, 4%), and full of energy (always, 25%; mostly, 54%; quite often, 16.7%; sometimes, 4.2%) after the procedure. Recipients of RSD indicated that they felt more light and healthy, and nearly all recipients (93%) described a loss of anxiety and indisposition. CONCLUSIONS: This investigation revealed that sufficient BP reduction by RSD and time following therapeutic success lead to significant improvements in patient QoL.

Image quality of ultra-low radiation exposure coronary CT angiography with an effective dose <0.1 mSv using high-pitch spiral acquisition and raw data-based iterative reconstruction.

OBJECTIVES: We evaluated the potential of prospectively ECG-triggered high-pitch spiral acquisition with low tube voltage and current in combination with iterative reconstruction to achieve coronary CT angiography with sufficient image quality at an effective dose below 0.1 mSv. METHODS: Contrast-enhanced coronary dual source CT angiography (2 × 128 × 0.6 mm, 80 kV, 50 mAs) in prospectively ECG-triggered high-pitch spiral acquisition mode was performed in 21 consecutive individuals (body weight <100 kg, heart rate ≤60/min). Images were reconstructed with raw data-based filtered back projection (FBP) and iterative reconstruction (IR). Image quality was assessed on a 4-point scale (1 = no artefacts, 4 = unevaluable). RESULTS: Mean effective dose was 0.06 ± 0.01 mSv. Image noise was significantly reduced in IR (128.9 ± 46.6 vs. 158.2 ± 44.7 HU). The mean image quality score was lower for IR (1.9 ± 1.1 vs. 2.2 ± 1.0, P < 0.0001). Of 292 coronary segments, 55 in FBP and 40 in IR (P = 0.12) were graded "unevaluable". In patients with a body weight ≤75 kg, both in FBP and in IR, the rates of fully evaluable segments were significantly higher in comparison to patients >75 kg. CONCLUSIONS: Coronary CT angiography with an estimated effective dose <0.1 mSv may provide sufficient image quality in selected patients through the combination of high-pitch spiral acquisition and raw data-based iterative reconstruction.